C4B and BAT2, assigned to the SLA class III region, were recently reported on relation with human diseases. The primers for RT-PCR and RACE-PCR for CDS analysis of these genes of pig were designed by aligning the CDSs of humans and mice from GenBank. After we amplified and sequenced with these primers and cDNAs, the full-length CDSs of pig were determined. The CDS lengths of C4B and BAT2 were shown as 5226 bp and 6501 bp. In addition, the identities of nucleotide sequences with human and mouse were 76% to 87%, and the identities of amino acids were 72% to 90%. After we carried out the alignment with determined CDSs in this study and pig genomic sequences from GenBank, the primers for cSNP detection in genome were designed in intron regions that flanked one or more exons. Then, we amplified and directly sequenced with genomic DNAs of six pig breeds. Four cSNPs from C4B and three 3 cSNPs from BAT2 were identified. In addition, amino acid substitution occurred in six cSNP positions except for C4248T of C4B. By the Multiplex-ARMS method, we genotyped seven cSNPs with DNA samples used for direct sequencing. We verified that this result was the same as that analyzed using direct sequencing. To demonstrate recrudescence, we performed both direct sequencing and Multiplex-ARMS on two randomly selected DNA samples. The genotype of each sample showed the same result from both methods. Therefore, seven cSNPs were identified from C4B and BAT2 and could be used as the basic data for haplotype analysis of SLA class III region. Moreover, the Multiplex-ARMS method should be powerful for genotyping of genes assigned to the whole SLA region for the xenograft study.

Recently, copy number variations (CNV) of genes or genomic segments have been intensively studied and various analysis methods have been developed. In this study, quantitative oligonucleotide ligation assay (qOLA) was applied to investigate CNV of KIT gene in the Landrace breed. A combined assay using qOLA and pyrosequencing, 6 genotype classes, I1/I1 or I3/i (IBe), I1/I2 or I3/IP, I1/I3, I1/IP or I2/i (IBe), I2/I2and I2/IP, were identified from 44 Landrace pigs. Genotype assignment using grouping features of measurements on a scatter plot showed 100% agreement with those using a statistical assignment by PROC FASTCLUS procedure implemented in the SAS package. Two versions (3100 and 3130) of ABI sequencers gave the same genotyping results, indicating there was no influence on qOLA by different versions of instrument, however, the means of standard deviation and coefficient of variation from the qOLA on a ABI 3130 (2.33 and 4.10) was lower than those from the qOLA on a ABI 3100 (2.67 and 4.81). Effect of proteinase K treatment on the PCR product followed by qOLA was very clear because noise peaks were disappeared and the observed ration fit better to the reference ratio corresponding to each genotype.

receptor(MC4R) gene and carcass traits was examined in randomly selected commercial pigs. A porcine MC4R gene was genotyped for Asp298Asn(nt. 892G>A) by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP). A total of three genotypes, A/A, A/G, and G/G, were found with 28.8, 22.8, and 48.4% frequencies, respectively. In the whole population, pigs containing 892A/- showed significantly higher marbling score than those of homozygotes G/G(P<0.05). Two homozygotes, A/A and G/G showed lower in meat color score but higher in water holding capacity than those of heterozygotes A/G(P<0.01). However, the carcass weight of the barrows containing wild type -/G was significantly higher(i.e. more than 2.5kg) than those of homozygotes A/A(P<0.05). The effects of each genotype on carcass traits in the gilts were similar to those of the whole population, but not in barrows, suggesting an unknown sex-related effect on carcass traits. This study suggested that the genotype MC4R A/- could improve the meat quality in the commercial pig production. However, since the genetic polymorphism of MC4R gene differentially affected the carcass traits in sex-related manner, therefore, both parameters, the sex and genotype, should be considered for marker-assisted selection in commercial pig production.

Adipocyte-specific secretory factor(ADSF)/resistin, an hormone, is a small cysteine-rich protein secreted from adipose tissue and ADSF/resistin has been implicated in modulating adipogenesis in human and rodents. Although the exact role of ADSF/resistin in bovine has not been identified, it may have directly or indirectly involved in adipocyte differentiation. The objective of this study was to investigate its DNA polymorphism associated with carcass traits in Korean Native Cattle(Hanwoo). To investigate DNA polymorphism in Hanwoo ADSF/resistin gene, blood samples were taken from 295 Hanwoo steers belonging to progeny testing at Hanwoo Improvement Center in Korea. Seven single nucleotide polymorphisms(SNPs) were found in intron regions but not in any other regions including promoter (1.7kb) and 4 exons. The highest frequency among SNPs was C186A(0.16/0.84) following G964A (0.156/0.884). The significant correlation(P<0.05) between the SNPs and economic traits was found on 764Ains associated with marbling but not from any other SNPs determined. A computer simulation was also conducted to assess the efficiency of marker assisted selection(MAS) versus the conventional breeding scheme. Results revealed that MAS was more efficient as a breeding tool compared to the conventional. In conclusion, ADSF/Resistin gene is one of candidate genes to evaluate the quality, especially marbling score, in Hanwoo.

This research was carried out in order to establish the production technique for Poong-san dog’s frozen semen, by examining the semen characteristic and the volume of glycerol added to the dilution solution, thawing temperature and sperm motility and viability as well as the motility using CASA according to time variation. Average semen volume was 5.9ml, sperm concentration 116.3×106 sperm/ml, total sperm number 789.3×106 sperm, motility 88.7±1.7% and viability 87.6±7.8%. When it was cryopreservation and thawed at different glycerol concentrated extender, it showed 52.7% motility and 57.7±10.3% viability at 7% glycerol, compared to other treatments. For semen cryogeny, at conditions of 5, 7cm and a height of 10cm for pre-cryogeny and maintaining the semen at 7cm from the surface of liquid nitrogen resulted in profitable motility and viability.

The current study was undertaken to determine the effects of sex steroid hormones(estrogen, testosterone and 19-nortestosterone) on differentiation and proliferation of pig preadipocytes. The preadipocytes were isolated from the backfat of new-born female pigs by collagenase digestion. 10－8M and 10－7M sex steroid hormones were treated to the cultured preadipocytes. Sex steroid hormones treated during the early stage of cell growth did not affect differentiation and proliferation of preadipocytes. However, testosterone and 19- nortestosterone treated during the late stage of cell growth stimulated differentiation of pig preadipocytes.

Effect of dietary krill meal levels on the cellular immunity was studied in broiler chicks activated immune response. One day old male broiler chicks(Ross) were fed the experimental krill meal 0.0(basal), 0.5, 1.0 and 2.0% diets for 3wks. Blood TNF-α activity, ovotransferrin level and Con A induced proliferation of PBMC and splenocytes after 24 hr(21 d age) of the croton oil 10㎕ injection intra- musculary at the age of 20 days compared to the control olive oil. Krill meal diets did not affect growth performance of broiler chicks and plasma ovotransferrin levels but decreased significantly(p<0.0001) TNF-α like activity and proliferation of PBMC relative to krill meal 0.0% diet. And the proliferation of splenocytes were significantly(p<0.05) increased in birds fed krill meal 1.0% diet relative to krill meal 0.5 and 2.0% diets. The croton oil injection induced a significant(p<0.0001) increases in the TNF-α activity or the PBMC proliferation and enhanced circulating ovotransferrin levels relative to the olive oil. In birds injected with the croton oil the proliferation of PBMC was reduced linearly with the increase of dietary krill meal levels, and the proliferation of splenocytes was decreased in the krill meal 1.0 and 2.0% diets relative to olive oil. These results indicated that dietary krill meal changed the innate and cellular immunity in broiler chicks activated by the injection of croton oil.

The current study was designed to define whether a blend of prunus mume extract(25%) containing lactic acid(75%) and grape seed extract(10ppm) could affect in vitro antimicrobial activity and growth performance, intestinal microflora, plasma biochemical profiles and digestive enzymes activities in broiler chickens. In paper disc agar diffusion test, we clearly observed antimicrobial activity against E. coli in response to prunus mume extract or a blend of prunus mume extract. For in vivo test, a total of ninety six 3-d-old male broiler chicks were assigned to basal diet(CON), basal diet supplemented with antibiotics (ANTI) and 0.5% a blend of prunus mume extract(PRNUS) until 35 days of age. Throughout the entire experimental period(3-35 days), there were no differences in BW and FCR between the birds fed the basal diet with antibiotics and the diet supplemented with a blend of prunus mume. However, ANTI group showed a significant increase in BW and total gain compared to CON group. The weights of digestive organs such as the pancreas and mucosal tissues were not affected by dietary treatments. There was no difference in plasma levels of glucose, cholesterol, AST and ALT activity. However, triglyceride in plasma increased(P<0.05) in the birds fed the diet supplemented with 0.5% a blend of prunus mume extract compared to those fed antibiotics supplemented diet. The activities of pancreatic trypsin and amylase, and intestinal hydrolase including disaccharidase were not affected by dietary treatment. The colony forming units(CFU) of lactobacillus in the lower ileal-cecum of the birds fed the diet supplemented with a blend of prunus mume extract was significantly(P<0.05) higher than that of birds fed antibiotic supplemented diet without affecting the CFU of E. coli. In conclusion, the birds fed the diet supplemented a blend of prunus mume as an alternative to antibiotics showed a similar growth performance and an significant increase in lactobacillus population compared with the birds fed basal and antibiotics supplemented diets.

The aim of this study was to isolate and characterize bacteriocin-producing bacteria from the intestine of duck to use as probiotics for livestock. A total of 416 strains were isolated from the small intestine and cecum of ducks and 13 isolates were finally selected after determinging inhibitory activity against pathogenic indicators by spot-on-lawn method. The selected strains were identified as Lactobacillus salivarius JWS 58, Lactobacillus plantarum JWS 1354, Pediococcus pentosaceus JWS 939, 7 strains of enterococci, and 3 strains of Escherichia coli. Lact. salivarius JWS 58, Ent. faecium JWS 833, and Ped. pentosaceus JWS 939 showed a strong inhibitory activity against Listeria monocytogenes. E. coli JWS 108 inhibited the growth of E. coli and Staphylococcus aureus. Lact. salivarius JWS 58 strain survived almost 50% in pH 2.5 phosphate buffer for 2 hr. Ped. pentosaceus JWS 939 and Lact. plantarum JWS 1354 showed strong amylolytic activity. These results suggest that a combination of bacteriocins or multispecies probiotics of the selected strains has a strong potential of alternative to antibiotics in livestock production.

This study was conducted to determine the protein requirement level in growing Jindo dog through nitrogen balance experiment. Twelve female dogs aged 18～20 weeks old were allotted one of 3 dietary treatments containing 21, 23 and 25% of crude protein. Average daily gain of dogs fed experimental diets containing 21, 23 and 25% of crude protein were 65.42, 79.58 and 99.17g/d, respectively, and there was a significant difference between 21 and 25% of crude protein treatments(p<0.05). Retained nitrogen were calculated 0.74, 0.96 and 1.31g/kgBW.75/d for dogs fed diets containing 21, 23 and 25% of crude protein, respectively, and were significantly higher(p<0.05) in dogs fed 25% of crude protein diet then those of other dogs. A quadratic regression equation was calculated between nitrogen intake(x) and nitrogen retention(y); y=-2.519x2+12.79x-14.79, and it was found a significantly(p<0.05) higher regression coefficient of 0.782. From the above equation, it was estimated maintenance requirement of crude protein for growing Jindo dog is 11.25g/kg BW.75/d.

To evaluate the effect of fiber sources on chewing activity, five sheep were consecutively fed diets containing 45% of a fiber source selected from 7 tested fiber sources of alfalfa hay cube(AHC), corn cob (CC), corn silage(CS), cotton seed hull(CSH), peanut hull(PHL), rice straw(RS) and sugarcane bagasse (SCB). Number of chew showed significantly higher value in CC(p<0.001) then other sources. RS and CC had highest(p<0.01) rumination times of 352 and 367 min/d, respectively. CC also showed the highest number of chew per kg NDF intake(p<0.01), but rumination time per kg NDF intake showed no difference except CSH(p<0.001). These results suggest that chewing activities were greatly affected by the fiber sources, and therefore it should be accounted in the formulation for ruminant feed.

In order to investigate the effects of supplemental ionic surfactants in in vitro ruminal fermentation, N-Lauroylsarcosine sodium salt(N-LSS) and sodium dodecyl sulfate(SDS) for negative(－) ionic surfactant, and hexadecylpyridinium chloride monohydrate(HPCM) and hexadecyltrimethyl ammonium bromide(HTAB) for positive (+) ionic surfactant were supplemented by 0.05% and 0.1% into the Dehority’s artificial medium containing rice straw(1mm) as a substrate. In vitro DM digestibility, the growth of rumen mixed microbes, pH, cumulative gas production and SEM(Scanning Electron Microscopy) observation of microbial attachment on rice straw particle were investigated through the experiment composing 9 treatments (two supplemental levels of two positive ionic(+) surfactant, two supplemental levels of two negative(－) ionic surfactant) including the control. The sample collection was at 6, 12, 24, 48 and 72 h post fermentation with 3 replications per treatments. DM digestibility in treatments supplemented (+) or (－) surfactants almost stopped afterward 12 h fermentation, in vitro DM digestibility at 72 h post fermentation in the ionic surfactants was at half level of that of the control(P<0.05). Accumulative gas production in in vitro was less(P<0.05) with addition of ionic surfactants compared to the control. The amount of rumen mixed microbes recovered from in vitro incubation fluid pleateaued at 12 h post fermentation for the positive (+) ionic surfactants, but steadily increased as fermentation time elapsed for the control. Rumen microbial growth rate was significantly(P<0.05) low in the negative(－) ionic surfactant compared to the control. pH of the incubation fluid was ranged from 6.02 to 7.20, and was the highest in the negative(－) ionic surfactants, and was the lowest in the control(P<0.05). In SEM observation, rumen microbial population attached on rice straw particle was less with addition of ionic surfactants than the control. In conclusion we could not found any positive effects of negative- and positive- charged surfactants on rumunal fermentation characteristics and rumen microbial growth rates.

This study was conducted to evaluate the effects of dietary addition of caprylic acid(CA)-cyclodextrin (CD) complex on in vitro fermentation characteristics, total gas and methane production. Experiment was done with six treatment groups; 1) no CA-CD complex(control), 2) CA 20 mg(T1), 3) CD 830 mg(T2), 4) CA-CD complex 425 mg(T3), CA-CD complex 850mg(T4), CA-CD complex 1,700 mg(T5). Ruminal pH, ammonia and total VFA concentrations of T2, T3, T4 and T5 were lower(P<0.05) than those of control and T1 for the 12h incubation. The increase in molar percentage of propionate was observed in T4 and T5 compared with control and T2 for the 8h incubation(P<0.05), however, the ratio of acetate to propionate was unchanged in all treatments. Total gas of T1 was lower than that of control, but T2, T3, T4 and T5 were higher compared with control for 12h incubation(P<0.05). If the methane ratio (as %) to total gas for all treatments was compared, T3, T4 and T5(CA-CD supplemented groups) averaged 2.7% whereas control, T1 and T2 showed 3.4, 2.8 and 5.1%, respectively. Therefore, according to these results, it might be concluded that supplementation of CA-CD complex could reduce methane production without disrupting ruminal fermentation.

This study was conducted to determine effects of cellulolytic microbes inoculation to sawdust-based spent mushroom substrate(SMS) during deepstacking on fermentation parameters, total microbial counts and cellulolytic enzyme activity and to on SMS nutrients utilization by sheep. For sheep metabolism trials, six sheep(ram, average 54.8kg) were fed a Control diet(70% concentrates, 15% rice straw and 15% SMS with no microbial treatment on a dry basis) and a Treatment diet(the same diet including SMS with a microbial treatment) for 2 trials. Spent mushroom substrates with or without a microbial(4 strains including 1 strain of Enterobacter ludwigii, 1 strain of Bacillus cereus and 2 strains of Bacillus subtillis) treatment (1% of SMS on wet basis) were deepstacked for 7 days. The internal temperatures in 1.2 M/T of SMS deepstacks reached to 50±5℃ within 7 days of storage. Total microbial counts remarkably decreased (P<0.05) with a deepstacking process and were not affected(P>0.05) by the microbial treatment. For fibrolytic enzyme activity, CMCase and xylanase activities were decreased(P<0.05) by a deepstacking process. After deepstacking, the microbial treatment showed about 2.5-times higher(P<0.05) for CMCase activity and about 4-times higher(P<0.05) for xylanase activity than those of the Control. Activities of ligninolytic enzymes such as laccase and MnP were not affected by the microbial treatment. The sheep fed the microbially treated SMS diet had a tendency of greater total tract digestibilities of ash(P=0.051), NFE (P=0.071), hemicellulose(P=0.087) and NDF(P=0.096) than those fed the untreated SMS diet. Nitrogen balance of sheep was not affected(P>0.05) by feeding of microbially treated SMS. Accordingly, these results indicate that cellulolytic microbes inoculation during deepstacking of SMS may improve the bio- utilization of SMS by sheep.